Kit for extracting DNA from histiocytes and method thereof

A tissue cell and kit technology, applied in the field of molecular biology, can solve the problems of affecting the experimental speed, the amount of DNA extracted is not high, and the time-consuming, etc., to achieve the effect of shortening the extraction time, high DNA content, and fast extraction speed.

Active Publication Date: 2011-01-05
中生方政生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the spin column method can guarantee the purity of the extracted DNA, the amount of extracted DNA is generally not high, which cannot meet the requirements of molecular biology experiments and clinical detection of nucleic acid levels.
In addition, the spin column method uses some toxic reagents, such as phenol, chloroform, etc., which is not conducive to the health of the experimenters; at the same time, the spin column method consumes a lot of time in the whole process of extracting DNA, which affects the speed of the next experiment

Method used

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  • Kit for extracting DNA from histiocytes and method thereof
  • Kit for extracting DNA from histiocytes and method thereof
  • Kit for extracting DNA from histiocytes and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1: kit of the present invention

[0081] The kit for extracting DNA from tissue cells of the present invention comprises:

[0082] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 . Specific preparation method: weigh 0.8gNaCl, 0.02g KCl, 0.024g KH respectively 2 PO 4 , 0.18g K 2 HPO 4 , add an appropriate amount of distilled water to dissolve and mix, then adjust the pH to 7.2 with HCl, and then adjust the volume to 100ml.

[0083] Lysis solution: 2M NaCl, pH8.020mM Tris-HCl, 25mM EDTA, 0.5% SDS. Specific preparation method: Measure 2ml of 1M Tris-HCl (pH 8.0), 5ml of 0.5M EDTA (pH 8.0), 40ml of 5M NaCl, and 5ml of 10% SDS, add appropriate amount of distilled water and adjust the volume to 100ml.

[0084] Binding solution: 4M guanidine isothiocyanate, pH 4.4 10mM Tris-HCl, 20% Ttiton-X 100, 10mM Urea. Specific preparation method: Weigh 47.264g guanidine isothiocyanate and 0.06g Urea respectively, add ap...

Embodiment 2

[0091] Example 2: DNA Extraction from Mouse Liver

[0092] DNA was extracted from mouse liver using the kit described in Example 1.

[0093] 1. Extraction reagent

[0094] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 .

[0095] Lysis solution: 2M NaCl, pH8.020mM Tris-HCl, 25mM EDTA, 0.5% SDS.

[0096] Binding solution: 4M guanidine isothiocyanate, pH4.4 10mM Tris-HCl, 20% Ttiton-X 100, 10mM Urea.

[0097] Conditioning binding solution: analytically pure isopropanol.

[0098] Biomagnetic beads: the core is Fe 3 o 4 , Peripheral wrapped silicon dioxide, product of Promega company.

[0099] Magnetic Separation Rack: Built-in magnets.

[0100] Protein removal solution: 5M guanidine hydrochloride, pH7.520mM Tris-HCl, 37% absolute ethanol.

[0101] Rinse solution: pH7.5 10mM Tris-HCl, 100mM NaCl, 80% absolute ethanol.

[0102] Eluent: Tris-HCl pH 8.0 10 mM.

[0103] 2. Extraction method

[0104] (1) Grind the liver tis...

Embodiment 3

[0121] Example 3: DNA extraction from pine needles

[0122] 1. The kit for extracting DNA from tissue cells according to the present invention

[0123] The kit for extracting DNA from tissue cells of the present invention comprises:

[0124] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 .

[0125] Lysis solution: 0.5M NaCl, pH6.55mM Tris-HCl, 10mM EDTA, 0.25% SDS.

[0126] Binding solution: 3M guanidine isothiocyanate, pH3.55mM Tris-HCl, 5% Ttiton-X100, 5mM Urea.

[0127] Conditioning binding solution: analytically pure isopropanol.

[0128] Biomagnetic beads: the core is Fe 3 o 4 , Peripheral wrapped silicon dioxide, product of Promega company.

[0129] Magnetic Separation Rack: Built-in magnets.

[0130] Protein removal solution: 2M guanidine hydrochloride, pH6.5 5mM Tris-HCl, 5% absolute ethanol.

[0131] Rinse solution: Tris-HCl at pH 6.55mM, NaCl at 50mM, and absolute ethanol at 50%.

[0132] Eluent: Tris-HCl, pH...

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Abstract

The invention relates to the field of molecular biology and discloses a kit and a method for extracting DNA from histiocytes. The kit comprises a binding solution, a regulating and binding solution and a deproteinization solution, wherein the binding solution comprises 3-5M of guanidinium isothiocyanate, 5-50mM of Tris-HCl with a pH value of 3.5-5.5, 5-25 percent by weight of Ttiton-X 100 and 5-20mM of Urea; the regulating and binding solution is isopropyl alcohol; and the deproteinization solution comprises 2-6M of guanidine hydrochloride, 5-50mM of Tris-HCl with a pH value of 6.5-8.0 and 5-50 percent by weight of absolute ethyl alcohol. The kit also comprises a pretreatment buffer solution, a splitting solution, a magnetic bead, a rinsing solution and an eluting solution. The DNA extracted by the kit and the method has the advantages of high content and high extraction speed, and the invention does not adopt toxic reagents and is suitable for extracting the DNA of various histiocytes.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for extracting DNA from tissue cells, and also to a method for extracting DNA from tissue cells. Background technique [0002] DNA, translated into Chinese as deoxyribonucleic acid, is the main component of chromosomes. DNA is the most important biological information molecule, which can form genetic instructions to guide the development of organisms and the operation of life functions. Transcribing genes and expressing proteins in an orderly manner in time and space completes all programs of directional development; preliminarily determines the unique traits and personalities of organisms and all stress responses when interacting with the environment. [0003] In addition to playing a very important role in the normal growth, development and reproduction of organisms and other life activities, it is also closely related to abnormal conditions of life, such as tumor occurr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 李艳萍
Owner 中生方政生物技术股份有限公司
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